Figure 2

miR-140 sequence and activity of rsmiR-140. A) Sequence of the miR-140 precursor (pre-miR-140) stem-loop located in intron 16 of the WWP2 gene. The letters in bold represent the seed regions of the mature miRNAs. The arrows delineate the DNA sequence upstream of the pre-miR-140 (shown between the diagonal lines) that was cloned into the pGL3-basic vector and identified as miR-140 regulatory sequence (rsmiR-140) with the location of potential transcription factor binding sites. B) The rsmiR-140 (1.153 kb and 1.883 kb) activities were determined in SW1353 cells (n = 4). Plasmid activity was calculated as relative luciferase units per μg of protein; the control (pGL3-basic) was assigned an arbitrary value of 1 for each experiment and the rsmiR-140 activity evaluated as fold change over control. P values were assessed by the one-sample t-test, comparing rsmiR-140 to the pGL3-basic vector.